Dramatic potentiation of the antiviral activity of HIV antibodies by cholesterol conjugation

Krzysztof Lacek, Richard A. Urbanowicz, Fulvia Troise, Claudia De Lorenzo, Valeria Severino, Antimo Di Maro, Alexander W. Tarr, Francesca Ferrara, Alexander Ploss, Nigel Temperton, Jonathan K. Ball, Alfredo Nicosia, Riccardo Cortese, Antonello Pessi

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11 Scopus citations

Abstract

The broadly neutralizing antibodies HIV 2F5 and 4E10, which bind to overlapping epitopes in the membrane-proximal external region of the fusion protein gp41, have been proposed to use a two-step mechanism for neutralization; first, they bind and preconcentrate at the viral membrane through their long, hydrophobic CDRH3 loops, and second, they form a high affinity complex with the protein epitope. Accordingly, mutagenesis of the CDRH3 can abolish their neutralizing activity, with no change in the affinity for the peptide epitope.Weshow here that we can mimic this mechanism by conjugating a cholesterol group outside of the paratope of an antibody. Cholesterol-conjugated antibodies bind to lipid raft domains on the membrane, and because of this enrichment, they show increased antiviral potency. In particular, we find that cholesterol conjugation (i) rescues the antiviral activity of CDRH3-mutated 2F5, (ii) increases the antiviral activity of WT 2F5, (iii) potentiates the non-membrane-binding HIV antibody D5 10-100-fold (depending on the virus strain), and (iv) increases synergy between 2F5 and D5. Conjugation can be made at several positions, including variable and constant domains. Cholesterol conjugation therefore appears to be a general strategy to boost the potency of antiviral antibodies, and, because membrane affinity is engineered outside of the antibody paratope, it can complement affinity maturation strategies.

Original languageEnglish (US)
Pages (from-to)35015-35028
Number of pages14
JournalJournal of Biological Chemistry
Volume289
Issue number50
DOIs
StatePublished - Dec 12 2014

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Lacek, K., Urbanowicz, R. A., Troise, F., De Lorenzo, C., Severino, V., Di Maro, A., Tarr, A. W., Ferrara, F., Ploss, A., Temperton, N., Ball, J. K., Nicosia, A., Cortese, R., & Pessi, A. (2014). Dramatic potentiation of the antiviral activity of HIV antibodies by cholesterol conjugation. Journal of Biological Chemistry, 289(50), 35015-35028. https://doi.org/10.1074/jbc.M114.591826